Stephen Snowdy <snowds at med.unc.edu> schrieb in im Newsbeitrag:
39A583F2.A8AFED6F at med.unc.edu...
> I fractionated cells by hypotonic buffer and dounce homogenization. On
> the advice (thank you for your help) of someone on this group, I used
> GADPH as a positive control
> for the cytoplasmic fraction. Should the GADPH show up only
> in the cyto fraction or will some also show up in the
> nuke fraction?
>> Once again, thank you for your help.
> --
> Stephen Snowdy
If it is in the nuclear fraction it can be regarded as impurity: GAPDH is a
glycolytic enzyme; glycolysis takes place in the cytoplasm. YOu could try to
further dilute your antibody or use less protein, the signals with this
antibody are usually very intense.
martin